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  • Title: Measurement of protein synthetic activity by determination of peptidyl[3H]puromycin formation in liver slices after ethanol administration.
    Author: Donohue TM, Sorrell JH, Sorrell MF, Tuma DJ.
    Journal: Alcohol Clin Exp Res; 1985 Dec; 9(6):526-30. PubMed ID: 3911813.
    Abstract:
    We investigated the utility of [3H]puromycin as an alternate and adjunct precursor to amino acids for measuring protein synthetic activity in rat liver slices. Slices were incubated in the presence of either [3H]puromycin or radiolabeled valine to compare the incorporation of these isotopic precursors into nascent hepatocellular proteins. Compared to liver slices from controls, comparable decreases in the incorporation of both [3H]puromycin and labeled valine were observed in experiments using slices from fasted rats and in slices preincubated with 25 mM ethanol. Radiolabeling of nascent polypeptides with either [3H]puromycin or labeled valine in liver slices from rats fed a liquid diet containing ethanol was also decreased compared to slices from pair-fed control and chow-fed animals. Our results demonstrated the validity of using [3H]puromycin to detect changes in protein synthetic activity under these conditions. The potential advantage of using [3H]puromycin for in vivo studies is discussed.
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